A new procedure has been developed for the preparation of radiolabeled chloramphenicol (CAP) and its stereoisomers. This method involves the reduction of CAP ketone with NaBH4 to quantitatively yield a 1:1 mixture of epimers, which are efficiently separated into the purified isomers by preparative HPLC. When NaBH4 is used as the reducing agent, this procedure can be used to synthesize CAP and its stereoisomers of high specific activity.